Fig. 1
Lentivirus vectors over-expressing mutant neurosin reduce the accumulation and propagation of α-synuclein. Wild-type neurosin (LV-wt-NR), neurosin-apoB (LV-wt-NR-apoB), the point mutant neurosin (LV-NR-R80Q) and R80Q-apoB (LV-NR-R80Q-apoB) were cloned into the 3rd generation lentivirus vector. a B103 neuronal cells were transduced with the vectors and examined by immunoblot for neurosin and the epitope tag V5. b B103 transduced cells were also immunostained for neurosin expression. c Compared to LV-Control, neuronal cells transduced with the neurosin constructs expressed higher levels of neurosin, levels were comparable between the wt and mutant neurosin constructs. d An in vitro neuronal co-culture system was devised to mimic the propagation of α-syn. B103 neuronal “Donor cells” infected with LV-α-syn or LV-Control (red) were plated in cell culture inserts containing a 0.4 μm membrane. B103 neuronal “Acceptor cells” infected with LV-NR-R80Q, LV-NR-R80Q-apoB or LV-Control were plated on coverslips. e Confocal microscopy and immunocytochemical analysis showing α-syn (red) from the Donor Cells was secreted and taken up by Acceptor cells that were double labeled with an antibody against neurosin (green). f Image analysis of double labeled neuronal cells, results are expressed as percent of acceptor cells containing α-syn immunoreactivity. * - indicates one way ANOVA with post hoc Dunnett’s, p < 0.05 compared to LV-Control. # - indicates one way ANOVA with post hoc Tukey-Krammer, p < 0.05 compared to LV-wt-NR. Scale bar = (B) 10 μm, (D) 5 μm